MicroRNA-135b promotes the proliferation of neuroblastoma cells by targeting forkhead box N3 and activating the phosphatidylinositol 3 kinase/protein kinase B signaling pathway
Objective To explore the effect of microRNA-135b(miR-135b)on the proliferation of neuroblastoma SH-SY5Y cells and its molecular mechanisms.Methods SH-SY5Y cells were transfected with miR-135b mimics,miR-135b inhibitor,and negative controls NC mimics,NC inhibitor,and grouped by simple randomization.The expression of miR-135b was detected by real-time quantitative polymerase chain reaction(RT-qPCR).Biological functions of miR-135b in migration,proliferation and invasion were examined by cell scratch,cell counting kit-8(CCK-8)and cell invasion assays.The bioinformatics soft-wares were used to predict target genes of miR-135b and luciferase reporter gene detection experiments were performed to verify it.The expression of phosphatidylinositol 3 kinase/protein kinase B(PI3K/Akt)signa-ling pathway and forhead box N3(FOXN3)protein was detected by Western blotting.T-test was used for inter group comparisons,and one-way ANOVA was used for inter group comparisons.Results The migra-tion,proliferation and invasion abilities in miR-135b mimics group were greater than those in NC mimics group[(28.47±1.10)％vs.(15.04±1.38)％,t=10.70,P＜0.01;1.97±0.11 vs.1.47±0.10,t=5.79,P＜0.05;265.00±21.93 vs.204.56±29.54,t=2.85,P＜0.05].The migration,prolifera-tion and invasion abilities in miR-135b inhibitor group were lower than those in NC inhibitor group[(7.67±1.24)％vs.(14.06±2.92)％,t=2.80,P＜0.05;1.04±0.08vs.1.49±0.12,t=5.39,P＜0.05;75.89±3.00 vs.201.11±19.78,t=10.84,P＜0.01].Bioinformatics softwares predicted FOXN3 was one of the target genes of miR-135b,which was confirmed by luciferase reporter assay.West-em blotting showed that FOXN3 expression was lower in the miR-135b mimics group than in the NC mimics group(0.35±0.15 vs.0.82±0.07,t=7.01,P＜0.05),and the expression of PI3K and p-Akt/Akt ra-tio were higher in the miR-135b mimics group than those in the NC mimics group(1.50±0.18 vs.0.85±0.05,t=4.97,P＜0.05;1.19±0.08 vs.0.89±0.02,t=5.54,P＜0.05);FOXN3 expression was higher in the miR-135b inhibitor group than in the NC inhibitor group(0.97±0.09 vs.0.75±0.10,t=3.87,P＜0.05),and the expression of PI3K and p-Akt/Akt ratio were lower in the miR-135b inhibitor group than those in the NC inhibitor group(0.77±0.11 vs.1.03±0.07,t=2.94,P＜0.05;0.64±0.01 vs.0.82±0.07,t=3.47,P＜0.05).Conclusion MiR-135b may target FOXN3 and activate PI3K/Akt signaling pathway to promote the proliferation of neuroblastoma cells.
MicroRNANeuroblastomaPhosphatidylinositol-3 kinase/protein kinase B signa-ling pathway
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